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You are provided with concentrated, purified, nucleic acid sample containing single DNA molecule_Describe how clectrophoresis could have been used to prepare this s...

Question

You are provided with concentrated, purified, nucleic acid sample containing single DNA molecule_Describe how clectrophoresis could have been used to prepare this sample and specify on factors affecting clectrophoretic mobility Could the size of the DNA molecule be estimated using this technique?(6) How would you go about determining if the DNA molecule was single-stranded or double- stranded? Be specific with regards to your proposed experiment(s) and the properties/data you are monitoring:(c)

You are provided with concentrated, purified, nucleic acid sample containing single DNA molecule_ Describe how clectrophoresis could have been used to prepare this sample and specify on factors affecting clectrophoretic mobility Could the size of the DNA molecule be estimated using this technique? (6) How would you go about determining if the DNA molecule was single-stranded or double- stranded? Be specific with regards to your proposed experiment(s) and the properties/data you are monitoring: (c) You have decided to perform digestion and obtained the following results: Enzyme Ecof [ Hpall HinaIII EcoRI Hpall EcoRI HindlHl Fragment Size (kb) 13,13 2.6 2.6 1.3,0.8,0.5 0.6,0.7,13 Is the original DNA molecule circular or linear? Prepare a map of the restriction enzyme sites indicating the distance between them;



Answers

A molecule of double-stranded DNA was cleaved with restriction nucleases, and the resulting products were separated by gel electrophoresis (Figure $Q 10-11$ ). DNA fragments of known sizes were electrophoresed on the same gel for use as size markers (left lane). The size of the DNA markers is given in kilobase pairs (kb), where $1 \mathrm{kb}=$ 1000 nucleotide pairs. Using the size markers as a guide, estimate the length of each restriction fragment obtained. From this information, construct a map of the original DNA molecule indicating the relative positions of all the restriction enzyme cleavage sites.

Our question is asking us what would happen once we use a promo. Um, immuno precipitation technique Thio Isolate a DNA fragment So we know that there are multiple forms of lab techniques used thio isolate a desired product. Um, you know, we waken die. There's different methods to isolate proteins. Thio isolate DNA, um, to die to isolate other, um, different molecules within the body. And so specifically this question is asking, how can we identify this DNA fragment? So one of the important things that was also covered in our textbook and in the chapter was it talked about poly A tails and it talked about, um It also talked about side A scene or CPG, right? So we talked about methylation. So when we're talking about controlling gene expression or even just isolating a fragment of DNA that we're interested in, maybe um manipulating to eventually activate or deactivate gene specific gene, we want toe, make sure that we get the right fragment, right? And that's what What? This question is acting so in the chapter. It also cover different post transcription allele methods. Andi, even other methods that air used to preserve DNA, and so that's what we do. We There's several different methods we can use. We can use a seat elation. We can use a little methylation or phosphor elation. So when we acetyl eight, what happens is in a Seattle group is added to the inter Minister of um ah, his stone protein. Um, and another post transcription aled modification that we can do is we can add poly a tails. And so the poly A tails are always going to be added on the three prime end. And this is to preserve or protect our, um, our DNA strand. This is very important. We're trying to preserve this fragment of DNA so that then we can eventually use it for, um, controlling different gene expressions. So we also have our five prime in right. We've got our five prime and three prime in. So generally what happens is the Poly A tails will be added on the three prime end and methylation or metal groups, so we'd add a CH three group on our five front end. This is also referred to as CPG s, and these air also protective molecules that are added to our five prime end of our DNA us fragment to preserve it and to keep it protected. And so when we're doing this immuno precipitation experiment, we want to tag these ends. So we know where our we know where our DNA fragment is. We know what's attached to it. So that then once we isolate it, we know that we have actually precipitated out, um, the correct molecule we were looking for. And so that's exactly what we're going to do is, um, tagged the this DNA fragment with a five crime or methylated and then add a poly, have a poly a tail, and therefore we have tagged our DNA fragment, and when we isolate it, we know that we'll have the right molecule.

Yeah, I feel this one is a plasma. So I'm gonna draw it as a circular cell. What to do? Big circle over here, We have a peek. All right, Okay, so starting here we have .6. Then we have bam you try and then we have .5 So little bit shorter, your hand three. And then between him and three and him three we've got to that's quite a bit longer. And then it's .7. No, we have. Then they try again And then after that is 2.7. So a really big gap meep again. This is not drawn to scale. Please keep that in mind. And then the last one is going to be the biggest. It's four.

Okay, so first let's create a little chart. So cora and kind three. Okay. So yeah, 2.9 killed c. Very called A plus B. Um and eight point oh equals B pussy. Okay. And then we have 3.9 equals a pussy. Six point oh equals two E. And then 12.9 Equals two b. plus defensa. So we would draw our map here. I'll draw out at the bottom. It's gonna be super duper long. So we would have in three here, Can you have your 2.9 um three points five and then a huge split before in three It's going to be the seven four fill pipes remind you. I am not drawing this to scale at all. This over here is to kill by its, I'll try to make that look a little bit closer. That's A C. O. R. I. And finally six kill the lights. So that's what it'll look like.

Okay so what I'm gonna do is draw the three and then connect them to each other. So this is gonna be the front E. Cora. Yes. Hey try and okay so here you're going to have he age E. So five and 3 and then that will be connected to to be each B. And that's going to be bye one six so that would connect to the other one. Because as you can see here these two are in common so that would probably connecting here and then we have E. C. O. Alright. Uh huh. Um H. One. And that's gonna be E. No here we will be and then E. Yeah So here it's four Here it's to be here at six and that will connect in here. Yeah.


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